Generation of a recombinant Newcastle disease virus expressing two foreign genes for use as a multivalent vaccine and gene therapy vector

Newcastle disease virus (NDV) has been used as a vector in the development of vaccines and gene therapy. A majority of these NDV vectors express only a single foreign gene through either an independent transcription unit (ITU) or an internal ribosomal entry site (IRES). In the present study, we combined the ITU and IRES methods to generate a novel NDV LaSota strain-based recombinant virus vectoring the red fluorescence protein (RFP) and the green fluorescence protein (GFP) genes. Biological assessments of the recombinant virus, rLS/IRES-RFP/GFP, showed that it was slightly attenuated in vivo, yet maintained similar growth dynamics and viral yields in vitro when compared to the parental LaSota virus. Expression of both the RFP and GFP was detected from the rLS/IRES-RFP/GFP virus-infected DF-1 cells by fluorescence microscopy. These data suggest that the rLS/IRES-RFP/GFP virus may be used as a multivalent vector for the development of vaccines and gene therapy agents.